Reduced brain UCP2 expression mediated by microRNA-503 contributes to increased stroke susceptibility in the high-salt fed stroke-prone spontaneously hypertensive rat

UCP2 maps nearby the lod score peak of STR1-stroke QTL in the SHRSP rat strain. We explored the potential contribution of UCP2 to the high-salt diet (JD)-dependent increased stroke susceptibility of SHRSP. Male SHRSP, SHRSR, two reciprocal SHRSR/SHRSP-STR1/QTL stroke congenic lines received JD for 4 weeks to detect brain UCP2 gene/protein modulation as compared with regular diet (RD). Brains were also analyzed for NF-κB protein expression, oxidative stress level and UCP2-targeted microRNAs expression level. Next, based on knowledge that fenofibrate and Brassica Oleracea (BO) stimulate UCP2 expression through PPARα activation, we monitored stroke occurrence in SHRSP receiving JD plus fenofibrate versus vehicle, JD plus BO juice versus BO juice plus PPARα inhibitor. Brain UCP2 expression was markedly reduced by JD in SHRSP and in the (SHRsr.SHRsp-(D1Rat134-Mt1pa)) congenic line, whereas NF-κB expression and oxidative stress level increased. The opposite phenomenon was observed in the SHRSR and in the (SHRsp.SHRsr-(D1Rat134-Mt1pa)) reciprocal congenic line. Interestingly, the UCP2-targeted rno-microRNA-503 was significantly upregulated in SHRSP and decreased in SHRSR upon JD, with consistent changes in the two reciprocal congenic lines. Both fenofibrate and BO significantly decreased brain microRNA-503 level, upregulated UCP2 expression and protected SHRSP from stroke occurrence. In vitro overexpression of microRNA-503 in endothelial cells suppressed UCP2 expression and led to a significant increase of cell mortality with decreased cell viability. Brain UCP2 downregulation is a determinant of increased stroke predisposition in high-salt-fed SHRSP. In this context, UCP2 can be modulated by both pharmacological and nutraceutical agents. The microRNA-503 significantly contributes to mediate brain UCP2 downregulation in JD-fed SHRSP

Protective Activity of Broccoli Sprout Juice in a Human Intestinal Cell Model of Gut Inflammation

Benefits to health from a high consumption of fruits and vegetables are well established and have been attributed to bioactive secondary metabolites present in edible plants. However, the effects of specific health-related phytochemicals within a complex food matrix are difficult to assess. In an attempt to address this problem, we have used elicitation to improve the nutraceutical content of seedlings of Brassica oleracea grown under controlled conditions. Analysis, by LC-MS, of the glucosinolate, isothiocyanate and phenolic compound content of juices obtained from sprouts indicated that elicitation induces an enrichment of several phenolics, particularly of the anthocyanin fraction. To test the biological activity of basal and enriched juices we took advantage of a recently developed in vitro model of inflamed human intestinal epithelium. Both sprouts’ juices protected intestinal barrier integrity in Caco-2 cells exposed to tumor necrosis factor under marginal zinc deprivation, with the enriched juice showing higher protection. Multivariate regression analysis indicated that the extent of rescue from stress-induced epithelial dysfunction correlated with the composition in bioactive molecules of the juices and, in particular, with a group of phenolic compounds, including several anthocyanins, quercetin-3-Glc, cryptochlorogenic, neochlorogenic and cinnamic acids

Negative Feedback Regulation of Auxin Signaling by ATHB8/ACL5–BUD2 Transcription Module

ABSTRACT The role of auxin as main regulator of vascular differentiation is well established, and a direct correlation between the rate of xylem differentiation and the amount of auxin reaching the (pro)cambial cells has been proposed. It has been suggested that thermospermine produced by ACAULIS5 (ACL5) and BUSHY AND DWARF2 (BUD2) is one of the factors downstream to auxin contributing to the regulation of this process in Arabidopsis . Here, we provide an in-depth characterization of the mechanism through which ACL5 modulates xylem differentiation. We show that an increased level of ACL5 slows down xylem differentiation by negatively affecting the expression of homeodomain-leucine zipper ( HD–ZIP ) III and key auxin signaling genes. This mechanism involves the positive regulation of thermospermine biosynthesis by the HD–ZIP III protein ARABIDOPSIS THALIANA HOMEOBOX8 tightly controlling the expression of ACL5 and BUD2 . In addition, we show that the HD–ZIP III protein REVOLUTA contributes to the increased leaf vascularization and long hypocotyl phenotype of acl5 likely by a direct regulation of auxin signaling genes such as LIKE AUXIN RESISTANT2 ( LAX2 ) and LAX3 . We propose that proper formation and differentiation of xylem depend on a balance between positive and negative feedback loops operating through HD–ZIP III genes

Glicosinolates in food

Glucosinolates are secondary plant metabolites that have attracted researcher’s attention due to their potential chemopreventive activity. More than 120 different glucosinolates have been identified in plants, and several of these compounds have been studied for the potential anti-cancerogenic effect of their metabolic breakdown products (mainly ITCs). Glucosinolates are peculiar of vegetables belonging to Brassicaceae family but are present also in few other species (capers, papaya, and moringa) used for human consumption. The type and concentration of glucosinolates in food are highly variable depending on several factors, such as genetics, cultivation site, cultivar, growth conditions, developmental stage, plant tissue, post-harvest handling, and food preparation methods. As types and concentration are also the main determinant of their biological activities, estimates of their content in food are essential tool to understand if a certain diet is adequate to deliver qualitatively and quantitatively appropriate glucosinolates and ITCs. The aim of this chapter is to describe qualitative and quantitative glucosinolate distribution among commonly eaten food, as well as the effect of the post-harvest handling on the glucosinolate food content

SCREENING GROWTH CONDITIONS TO INCREASE ANTHOCYANINS AND GLUCOSINOLATES CONTENT IN BROCCOLI SPROUTS

Broccoli (Brassicaceae) are an excellent dietary source of micronutrients (vitamins and minerals) and health-promoting phytochemicals (glucosinolates and phenolic compounds). Glucosinolates are a class of organic compounds derived from glucose and different aminoacids, and characterized by the high number of different side chains. Anthocyanins represent the largest group of natural, water-soluble plant pigments, and approximately 400 anthocyanins have been determined. Glucosinolates and anthocyanins protects plants against various biotic and abiotic stresses and possess antioxidant and antinflammatory activity in humans. In addition, glucosinolates have gained much attention due to the anticarcinogenic activity of isothiocyanates, their major hydrolysis products, molecules able to induce phase 2 detoxication enzymes and protect animals against chemically induced cancer (Zhang, 2012). Young broccoli plant are an especially good source of glucosinolates and anthocyanins and several studies report that growth and environmental conditions could affect the profiles of these secondary plant metabolites (Perez-Balibrea et al., 2008). We previously developed a rapid and sensitive LC-MS (MRM) method for direct and simultaneous quantification of 14 glucosinolates in broccoli sprouts. In this study, the effect of different abiotic stresses and hormones involved in plant defence responses of the accumulation of glucosinolates and anthocyanins in broccoli sprouts was investigated. Quantitative results show that all treatments modify glucosinolates and anthocyanins profile. In particular, light grown broccoli sprouts treated with sucrose show glucosinolates and anthocyanins levels double than control. This work was ,supported by Italian Ministry of Agriculture, Food & Forestry (MiPAAF) grant "NUTRIGEA" (DM 30281 23112/2009)

A simple microsatellite-based method for hazelnut oil DNA analysis

Molecular food traceability requires continuous updates to identify more robust, efficient and affordable methodologies to guarantee food quality and safety and especially consumers’ health. Available commercial kits are often unsatisfactory and require modifications to successfully detect single components on complex and transformed food matrices. Here we report a simple method for molecular traceability of cold-pressed hazelnut oil based on microsatellite DNA markers. Different genomic extraction methodologies were tested and a total genome pre-amplification step was applied on PCR-negative samples. PCR-capillary electrophoresis using nine microsatellites demonstrates the accuracy of the fingerprint analysis even for filtered oil

Untargeted Metabolomics Reveals Predominant Alterations in Lipid Metabolism Following Light Exposure in Broccoli Sprouts

The consumption of vegetables belonging to the family Brassicaceae (e.g., broccoli and cauliflower) is linked to a reduced incidence of cancer and cardiovascular diseases. The molecular composition of such plants is strongly affected by growing conditions. Here we developed an unbiased metabolomics approach to investigate the effect of light and dark exposure on the metabolome of broccoli sprouts and we applied such an approach to provide a bird’s-eye view of the overall metabolic response after light exposure. Broccoli seeds were germinated and grown hydroponically for five days in total darkness or with a light/dark photoperiod (16 h light/8 h dark cycle). We used an ultra-performance liquid-chromatography system coupled to an ion-mobility, time-of-flight mass spectrometer to profile the large array of metabolites present in the sprouts. Differences at the metabolite level between groups were analyzed using multivariate statistical analyses, including principal component analysis and correlation analysis. Altered metabolites were identified by searching publicly available and in-house databases. Metabolite pathway analyses were used to support the identification of subtle but significant changes among groups of related metabolites that may have gone unnoticed with conventional approaches. Besides the chlorophyll pathway, light exposure activated the biosynthesis and metabolism of sterol lipids, prenol lipids, and polyunsaturated lipids, which are essential for the photosynthetic machinery. Our results also revealed that light exposure increased the levels of polyketides, including flavonoids, and oxylipins, which play essential roles in the plant’s developmental processes and defense mechanism against herbivores. This study highlights the significant contribution of light exposure to the ultimate metabolic phenotype, which might affect the cellular physiology and nutritional value of broccoli sprouts. Furthermore, this study highlights the potential of an unbiased omics approach for the comprehensive study of the metabolism

The Arabidopsis ATHB-8 HD-Zip Protein Acts as a Differentiation-Promoting Transcription Factor of the Vascular Meristems

ATHB-8, -9, -14, -15, and IFL1/REV are members of a small homeodomain-leucine zipper family whose genes are characterized by expression in the vascular tissue. ATHB-8, a gene positively regulated by auxin (Baima et al., 1995), is considered an early marker of the procambial cells and of the cambium during vascular regeneration after wounding. Here, we demonstrate that although the formation of the vascular system is not affected in athb8 mutants, ectopic expression of ATHB-8 in Arabidopsis plants increased the production of xylem tissue. In particular, a careful anatomical analysis of the transgenic plants indicated that the overexpression of ATHB-8 promotes vascular cell differentiation. First, the procambial cells differentiated precociously into primary xylem. In addition, interfascicular cells also differentiated precociously into fibers. Finally, the transition to secondary growth, mainly producing xylem, was anticipated in transgenic inflorescence stems compared with controls. The stimulation of primary and secondary vascular cell differentiation resulted in complex modifications of the growth and development of the ATHB-8 transgenic plants. Taken together, these results are consistent with the hypothesis that ATHB-8 is a positive regulator of proliferation and differentiation, and participates in a positive feedback loop in which auxin signaling induces the expression of ATHB-8, which in turn positively modulates the activity of procambial and cambial cells to differentiate

Improvement of the nutraceutical quality of broccoli sprouts by elicitation

Epidemiological studies show an inverse association between Brassica consumption and chronic diseases. Phytochemicals are thought to be beneficial for human health and therefore responsible for this protective effect. Increasing their levels into Brassica food is considered an expedient nutritional strategy that can be achieved through the manipulation of growth conditions by elicitors. In this work we systematically evaluated the influence of treatment with different elicitors (sucrose, mannitol, NaCl, 1-aminocyclopropane-l-carboxylic acid, salicylic acid and methyl jasmonate) on the phytochemical composition of broccoli sprouts. The content of total and single glucosinolates, total phenolic compounds, total flavonoids, total anthocyanins, vitamin C and E and β-carotene was assessed. The exposure to different elicitors produced concentration- and elicitor-dependent specific changes in the content of all the phytochemicals considered. Sucrose, identified as the most effective elicitor by principal component analysis, induced a significant increase of total and specific glucosinolates, vitamin C, total anthocyanins and polyphenols. Sucrose is likely to represent an effective tool to increase the nutritional value of broccoli sprouts

Neuroprotective Effect of Brassica oleracea Sprouts Crude Juice in a Cellular Model of Alzheimer’s Disease

β-Amyloid peptide (Aβ) aberrant production and aggregation are major factors implicated in the pathogenesis of Alzheimer’s disease (AD), causing neuronal death via oxidative stress. Several studies have highlighted the importance of polyphenolic antioxidant compounds in the treatment of AD, but complex food matrices, characterized by a different relative content of these phytochemicals, have been neglected. In the present study, we analyzed the protective effect on SH-SY5Y cells treated with the fragment Aβ25–35 by two crude juices of broccoli sprouts containing different amounts of phenolic compounds as a result of different growth conditions. Both juices protected against Aβ-induced cytotoxicity and apoptotic cell death as evidenced by cell viability, nuclear chromatin condensation, and apoptotic body formation measurements. These effects were mediated by the modulation of the mitochondrial function and of the HSP70 gene transcription and expression. Furthermore, the juices upregulated the intracellular glutathione content and mRNA levels or activity of antioxidant enzymes such as heme oxygenase-1, thioredoxin, thioredoxin reductase, and NAD(P)H:quinone oxidoreductase 1 via activation of NF-E2-related factor 2 (Nrf2). Although the effects of the two juices were similar, the juice enriched in phenolic compounds showed a greater efficacy in inducing the activation of the Nrf2 signalling pathway